腫瘤細胞需要透過改變他們的代謝路徑才能快速生長，其中丙酮酸激酶PKM2 (Pyruvate Kinase M2)扮演著重要的角色。PKM2是調控細胞糖解作用的關鍵酵素，它會使磷酸烯醇式丙酮酸轉化為丙酮酸。在腫瘤中，癌細胞內主要存在著低活性的二聚體PKM2使氧化磷酸化被抑制，留住糖解作用產生的中間代謝物做為DNA及蛋白質的基底以促進細胞快速增生。二聚體PKM2同時也會轉入核中去活化其下游基因的表現。在這篇論文中，我們確定了硫化氫會透過硫化蛋白質的後修飾調節PKM2的活性：硫化氫會將PKM2上的半胱氨酸硫化使四聚體PKM2被解離為低活性的二聚體或單體，而我們進一步找出PKM2上會被硫化的位點，並驗證該位點的胺基酸突變後可成功地阻斷PKM2的硫化，並且使其產生高活性的PKM2四聚體。細胞表達此突變後可以成功恢復PKM2的活性且阻止其入核，並增加活性氧類ROS（Reactive oxygen species）的產生從而抑制細胞增生及腫瘤的生長。總結以上，我們的研究顯示PKM2的硫化後修飾是癌症發展時改變代謝路徑的一種全新機制，使用此新穎代謝途徑來治療癌症可能在未來有一定的潛力。

Tumor cells require to grow rapidly by altering their metabolic pathways, in which pyruvate kinase M2 (PKM2) plays a critical role. PKM2 is the key enzyme in regulating glucose metabolism, which converts phosphoenolpyruvate (PEP) to pyruvate. In cancer cells, the less-active dimeric PKM2 is generated to suppress oxidative phosphorylation, resulting in the accumulation of glycolytic metabolites as building materials to accelerate cell division. The dimeric PKM2 is then translocated into the nucleus to activate gene expressions. Here we demonstrate that hydrogen sulfide (H2S) modulates PKM2 activity through a novel post-translational modification, protein sulfhydration. PKM2 tetramers dissociate into dimers or monomers with low enzyme activity through H2S mediated protein sulfhydration. Single amino acid mutation on the sulfhydrated site successfully blocks PKM2 sulfhydration, thereby promoting the formation of active tetramer forms of PKM2. Restoring PKM2 activity by expressing this mutant leads to the increased reactive oxygen species (ROS), which in turn inhibits cell proliferation and tumor growth. Overall, our study reveals that PKM2 sulfhydration serves as a novel mechanism for metabolic reprogramming during cancer development and may have therapeutic potentials in treating cancer specific metabolism.

摘要-----i
Abstract-----ii
致謝-----iii
Table of Contents-----iv
Table of Figures-----vi
Introduction-----1
1.1 Cancer Metabolism-----1
1.2 Pyruvate Kinase-----2
1.3 Regulation of Pyruvate Kinase M2 Activity-----3
Motivation and Aims-----8
3.1 Cell culture and transfection-----10
3.2 Generating stable cell line with different types of PKM2-----11
3.3 Immunoblotting-----12
3.4 Biotin switch (S-sulfhydration) assay-----12
3.5 Pyruvate kinase activities-----13
3.6 Glycerol gradient ultracentrifugation-----13
3.7 Confocal microscopic analysis-----14
3.8 ROS Measurement-----15
3.9 Proliferation assay-----16
Results-----18
4.1 Hydrogen sulfide promotes sulfhydration on PKM2-----18
4.2 PKM2 sulfhydration promotes the nuclear translocation of PKM2-----18
4.3 Establishment of cell lines stably expressing PKM2 wild type and mutant form.-----19
4.4 Blocking PKM2 sulfhydration increases pyruvate kinase activity-----20
4.5 PKM2 sulfhydration induces subunit dissociation from tetramer to dimer forms-----20
4.6 Blockage of sulfhydration site reduces PKM2 nuclear translocation-----21
4.7 Blockage of PKM2 sulfhydration induces production of reactive oxygen species (ROS)-----21
4.8 PKM2 sulfhydration is required for cancer cell proliferation and tumor growth.-----22
Conclusion-----24
Discussion-----26
Figures-----30
References-----43

[1] G.W. Prager, S. Braga, B. Bystricky, C. Qvortrup, C. Criscitiello, E. Esin, G.S. Sonke, G.A. Martinez, J.S. Frenel, M. Karamouzis, M. Strijbos, O. Yazici, P. Bossi, S. Banerjee, T. Troiani, A. Eniu, F. Ciardiello, J. Tabernero, C.C. Zielinski, P.G. Casali, F. Cardoso, J.Y. Douillard, S. Jezdic, K. McGregor, G. Bricalli, M. Vyas, A. Ilbawi, Global cancer control: responding to the growing burden, rising costs and inequalities in access, ESMO Open 3 (2018) e000285.
[2] W.H. Organization, WHO report on cancer: setting priorities, investing wisely and providing care for all, (2020).
[3] O.J.S. Warburg, On the origin of cancer cells, Science 123 (1956) 309-314.
[4] V.R. Fantin, J. St-Pierre, P.J.C.c. Leder, Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance, Cancer Cell 9 (2006) 425-434.
[5] B. Altenberg, K.O. Greulich, Genes of glycolysis are ubiquitously overexpressed in 24 cancer classes, Genomics 84 (2004) 1014-1020.
[6] P.K. Majumder, P.G. Febbo, R. Bikoff, R. Berger, Q. Xue, L.M. McMahon, J. Manola, J. Brugarolas, T.J. McDonnell, T.R.J.N.m. Golub, mTOR inhibition reverses Akt-dependent prostate intraepithelial neoplasia through regulation of apoptotic and HIF-1-dependent pathways, Nat. Med. 10 (2004) 594-601.
[7] K. Imamura, T. Tanaka, [25] Pyruvate kinase isozymes from rat, Methods in enzymology, Elsevier1982, pp. 150-165.
[8] M.S. Jurica, A. Mesecar, P.J. Heath, W. Shi, T. Nowak, B.L. Stoddard, The allosteric regulation of pyruvate kinase by fructose-1,6-bisphosphate, Structure 6 (1998) 195-210.
[9] H.R. Christofk, M.G. Vander Heiden, M.H. Harris, A. Ramanathan, R.E. Gerszten, R. Wei, M.D. Fleming, S.L. Schreiber, L.C.J.N. Cantley, The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth, Nature 452 (2008) 230-233.
[10] Q. Sun, X. Chen, J. Ma, H. Peng, F. Wang, X. Zha, Y. Wang, Y. Jing, H. Yang, R.J.P.o.t.N.A.o.S. Chen, Mammalian target of rapamycin up-regulation of pyruvate kinase isoenzyme type M2 is critical for aerobic glycolysis and tumor growth, Proc. Natl. Acad. Sci. U. S. A. 108 (2011) 4129-4134.
[11] D. Anastasiou, Y. Yu, W.J. Israelsen, J.K. Jiang, M.B. Boxer, B.S. Hong, W. Tempel, S. Dimov, M. Shen, A. Jha, H. Yang, K.R. Mattaini, C.M. Metallo, B.P. Fiske, K.D. Courtney, S. Malstrom, T.M. Khan, C. Kung, A.P. Skoumbourdis, H. Veith, N. Southall, M.J. Walsh, K.R. Brimacombe, W. Leister, S.Y. Lunt, Z.R. Johnson, K.E. Yen, K. Kunii, S.M. Davidson, H.R. Christofk, C.P. Austin, J. Inglese, M.H. Harris, J.M. Asara, G. Stephanopoulos, F.G. Salituro, S. Jin, L. Dang, D.S. Auld, H.W. Park, L.C. Cantley, C.J. Thomas, M.G. Vander Heiden, Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis, Nat. Chem. Biol. 8 (2012) 839-847.
[12] S.Y. Lunt, M.G. Vander Heiden, Aerobic glycolysis: meeting the metabolic requirements of cell proliferation, Annu Rev Cell Dev Biol 27 (2011) 441-464.
[13] W. Luo, H. Hu, R. Chang, J. Zhong, M. Knabel, R. O'Meally, R.N. Cole, A. Pandey, G.L.J.C. Semenza, Pyruvate kinase M2 is a PHD3-stimulated coactivator for hypoxia-inducible factor 1, Cell 145 (2011) 732-744.
[14] L.J. Reitzer, B.M. Wice, D.J.J.o.B.C. Kennell, Evidence that glutamine, not sugar, is the major energy source for cultured HeLa cells, J. Biol. Chem. 254 (1979) 2669-2676.
[15] K. Bluemlein, N.-M. Grüning, R.G. Feichtinger, H. Lehrach, B. Kofler, M.J.O. Ralser, No evidence for a shift in pyruvate kinase PKM1 to PKM2 expression during tumorigenesis, Oncotarget 2 (2011) 393.
[16] D.Y. Gui, C.A. Lewis, M.G. Vander Heiden, Allosteric regulation of PKM2 allows cellular adaptation to different physiological states, Sci Signal 6 (2013) pe7.
[17] Q. Xu, L. Liu, Y. Yin, J. He, Q. Li, X. Qian, Y. You, Z. Lu, S. Peiper, Y.J.O. Shu, Regulatory circuit of PKM2/NF-κB/miR-148a/152-modulated tumor angiogenesis and cancer progression, Oncogene 34 (2015) 5482-5493.
[18] R.A. Cairns, I.S. Harris, T.W. Mak, Regulation of cancer cell metabolism, Nat Rev Cancer 11 (2011) 85-95.
[19] J.D. Dombrauckas, B.D. Santarsiero, A.D. Mesecar, Structural basis for tumor pyruvate kinase M2 allosteric regulation and catalysis, Biochemistry 44 (2005) 9417-9429.
[20] S. Mazurek, Pyruvate kinase type M2: a key regulator within the tumour metabolome and a tool for metabolic profiling of tumours, Oncogenes Meet Metabolism, Springer2008, pp. 99-124.
[21] E. Eigenbrodt, M. Reinacher, U. Scheefers-Borchel, H. Scheefers, R.J.C.r.i.o. Friis, Double role for pyruvate kinase type M2 in the expansion of phosphometabolite pools found in tumor cells, Crit Rev Oncog. 3 (1992) 91.
[22] S. Mazurek, Pyruvate kinase type M2: a key regulator of the metabolic budget system in tumor cells, Int J Biochem Cell Biol 43 (2011) 969-980.
[23] S. Mazurek, C.B. Boschek, F. Hugo, E. Eigenbrodt, Pyruvate kinase type M2 and its role in tumor growth and spreading, Seminars in cancer biology, Elsevier, 2005, pp. 300-308.
[24] H.R. Christofk, M.G. Vander Heiden, M.H. Harris, A. Ramanathan, R.E. Gerszten, R. Wei, M.D. Fleming, S.L. Schreiber, L.C. Cantley, The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth, Nature 452 (2008) 230-U274.
[25] P. Presek, M. Reinacher, E.J.F.l. Eigenbrodt, Pyruvate kinase type M2 is phosphorylated at tyrosine residues in cells transformed by Rous sarcoma virus, FEBS Lett. 242 (1988) 194-198.
[26] L. Lv, D. Li, D. Zhao, R. Lin, Y. Chu, H. Zhang, Z. Zha, Y. Liu, Z. Li, Y.J.M.c. Xu, Acetylation targets the M2 isoform of pyruvate kinase for degradation through chaperone-mediated autophagy and promotes tumor growth, Mol. Cell 42 (2011) 719-730.
[27] S.H. Park, O. Ozden, G. Liu, H.Y. Song, Y. Zhu, Y. Yan, X. Zou, H.J. Kang, H. Jiang, D.R. Principe, Y.I. Cha, M. Roh, A. Vassilopoulos, D. Gius, SIRT2-Mediated Deacetylation and Tetramerization of Pyruvate Kinase Directs Glycolysis and Tumor Growth, Cancer Res 76 (2016) 3802-3812.
[28] G. Prakasam, M.A. Iqbal, R.N. Bamezai, S.J.F.i.o. Mazurek, Posttranslational modifications of pyruvate kinase M2: tweaks that benefit cancer, Front. Oncol. 8 (2018) 22.
[29] E.K. Wiese, T. Hitosugi, Tyrosine Kinase Signaling in Cancer Metabolism: PKM2 Paradox in the Warburg Effect, Front Cell Dev Biol 6 (2018) 79.
[30] R. Ruiter, L.E. Visser, M.P. van Herk-Sukel, J.-W.W. Coebergh, H.R. Haak, P.H. Geelhoed-Duijvestijn, S.M. Straus, R.M. Herings, B.H.C.J.D.c. Stricker, Lower risk of cancer in patients on metformin in comparison with those on sulfonylurea derivatives: results from a large population-based follow-up study, Diabetes Care 35 (2012) 119-124.
[31] S. Dalva-Aydemir, R. Bajpai, M. Martinez, K.U. Adekola, I. Kandela, C. Wei, S. Singhal, J.E. Koblinski, N.S. Raje, S.T. Rosen, M. Shanmugam, Targeting the metabolic plasticity of multiple myeloma with FDA-approved ritonavir and metformin, Clin Cancer Res 21 (2015) 1161-1171.
[32] A. Silvestri, F. Palumbo, I. Rasi, D. Posca, T. Pavlidou, S. Paoluzi, L. Castagnoli, G.J.P.o. Cesareni, Metformin induces apoptosis and downregulates pyruvate kinase M2 in breast cancer cells only when grown in nutrient-poor conditions, PLoS One 10 (2015) e0136250.
[33] D.R. Johnson, B.P.J.J.o.n.-o. O’Neill, Glioblastoma survival in the United States before and during the temozolomide era, J. Neurooncol. 107 (2012) 359-364.
[34] I. Park, J. Mukherjee, M. Ito, M.M. Chaumeil, L.E. Jalbert, K. Gaensler, S.M. Ronen, S.J. Nelson, R.O.J.C.r. Pieper, Changes in pyruvate metabolism detected by magnetic resonance imaging are linked to DNA damage and serve as a sensor of temozolomide response in glioblastoma cells, Cancer Res. 74 (2014) 7115-7124.
[35] C. Chen, W. Xiao, L. Huang, G. Yu, J. Ni, L. Yang, R. Wan, G. Hu, Shikonin induces apoptosis and necroptosis in pancreatic cancer via regulating the expression of RIP1/RIP3 and synergizes the activity of gemcitabine, Am J Transl Res 9 (2017) 5507-5517.
[36] T.-J. Lin, H.-T. Lin, W.-T. Chang, P.-W. Hsiao, S.-Y. Yin, N.-S.J.M.c. Yang, Shikonin-enhanced cell immunogenicity of tumor vaccine is mediated by the differential effects of DAMP components, Mol. Cancer 14 (2015) 1-13.
[37] J. Chen, J. Xie, Z. Jiang, B. Wang, Y. Wang, X. Hu, Shikonin and its analogs inhibit cancer cell glycolysis by targeting tumor pyruvate kinase-M2, Oncogene 30 (2011) 4297-4306.
[38] J.C. Tang, J. Zhao, F. Long, J.Y. Chen, B. Mu, Z. Jiang, Y. Ren, J. Yang, Efficacy of Shikonin against Esophageal Cancer Cells and its possible mechanisms in vitro and in vivo, J Cancer 9 (2018) 32-40.
[39] T. Tao, Q. Su, S. Xu, J. Deng, S. Zhou, Y. Zhuang, Y. Huang, C. He, S. He, M.J.J.o.C.P. Peng, Down‐regulation of PKM2 decreases FASN expression in bladder cancer cells through AKT/mTOR/SREBP‐1c axis, J. Cell. Physiol. 234 (2019) 3088-3104.
[40] J.C. Boulos, M. Rahama, M.F. Hegazy, T. Efferth, Shikonin derivatives for cancer prevention and therapy, Cancer Lett 459 (2019) 248-267.
[41] G. Yang, L. Wu, B. Jiang, W. Yang, J. Qi, K. Cao, Q. Meng, A.K. Mustafa, W. Mu, S.J.S. Zhang, H2S as a physiologic vasorelaxant: hypertension in mice with deletion of cystathionine γ-lyase, Science 322 (2008) 587-590.
[42] C. Coletta, A. Papapetropoulos, K. Erdelyi, G. Olah, K. Modis, P. Panopoulos, A. Asimakopoulou, D. Gero, I. Sharina, E. Martin, C. Szabo, Hydrogen sulfide and nitric oxide are mutually dependent in the regulation of angiogenesis and endothelium-dependent vasorelaxation, Proc Natl Acad Sci U S A 109 (2012) 9161-9166.
[43] R.C. Zanardo, V. Brancaleone, E. Distrutti, S. Fiorucci, G. Cirino, J.L. Wallace, R.C. Zanardo, V. Brancaleone, E. Distrutti, S.J.T.F.j. Fiorucci, Hydrogen sulfide is an endogenous modulator of leukocyte‐mediated inflammation, The FASEB journal 20 (2006) 2118-2120.
[44] X. Chen, K.H. Jhee, W.D. Kruger, Production of the neuromodulator H2S by cystathionine beta-synthase via the condensation of cysteine and homocysteine, J. Biol. Chem. 279 (2004) 52082-52086.
[45] B.D. Paul, S.H.J.N.r.M.c.b. Snyder, H 2 S signalling through protein sulfhydration and beyond, Nature reviews Molecular cell biology 13 (2012) 499-507.
[46] A.K. Mustafa, G. Sikka, S.K. Gazi, J. Steppan, S.M. Jung, A.K. Bhunia, V.M. Barodka, F.K. Gazi, R.K. Barrow, R.J.C.r. Wang, Hydrogen sulfide as endothelium-derived hyperpolarizing factor sulfhydrates potassium channels, Circ. Res. 109 (2011) 1259-1268.
[47] N. Krishnan, C. Fu, D.J. Pappin, N.K. Tonks, H2S-Induced sulfhydration of the phosphatase PTP1B and its role in the endoplasmic reticulum stress response, Sci Signal 4 (2011) ra86.
[48] N. Sen, B.D. Paul, M.M. Gadalla, A.K. Mustafa, T. Sen, R. Xu, S. Kim, S.H.J.M.c. Snyder, Hydrogen sulfide-linked sulfhydration of NF-κB mediates its antiapoptotic actions, Mol. Cell 45 (2012) 13-24.
[49] K. Módis, P. Panopoulos, C. Coletta, A. Papapetropoulos, C.J.B.P. Szabo, Hydrogen sulfide-mediated stimulation of mitochondrial electron transport involves inhibition of the mitochondrial phosphodiesterase 2A, elevation of cAMP and activation of protein kinase A, Biochem. Pharmacol. 86 (2013) 1311-1319.
[50] K. Módis, Y. Ju, A. Ahmad, A.A. Untereiner, Z. Altaany, L. Wu, C. Szabo, R.J.P.r. Wang, S-Sulfhydration of ATP synthase by hydrogen sulfide stimulates mitochondrial bioenergetics, Pharmacol. Res. 113 (2016) 116-124.
[51] A.A. Untereiner, G. Olah, K. Modis, M.R. Hellmich, C. Szabo, H2S-induced S-sulfhydration of lactate dehydrogenase a (LDHA) stimulates cellular bioenergetics in HCT116 colon cancer cells, Biochem Pharmacol 136 (2017) 86-98.
[52] M. Liang, S. Jin, D.D. Wu, M.J. Wang, Y.C. Zhu, Hydrogen sulfide improves glucose metabolism and prevents hypertrophy in cardiomyocytes, Nitric Oxide 46 (2015) 114-122.
[53] A. Papapetropoulos, A. Pyriochou, Z. Altaany, G. Yang, A. Marazioti, Z. Zhou, M.G. Jeschke, L.K. Branski, D.N. Herndon, R.J.P.o.t.N.A.o.S. Wang, Hydrogen sulfide is an endogenous stimulator of angiogenesis, Proc. Natl. Acad. Sci. U. S. A. 106 (2009) 21972-21977.
[54] W.J. Cai, M.J. Wang, L.H. Ju, C. Wang, Y.C. Zhu, Hydrogen sulfide induces human colon cancer cell proliferation: role of Akt, ERK and p21, Cell Biol Int 34 (2010) 565-572.
[55] Z. Ma, Q. Bi, Y. Wang, Hydrogen sulfide accelerates cell cycle progression in oral squamous cell carcinoma cell lines, Oral Dis 21 (2015) 156-162.
[56] Y.H. Wang, J.T. Huang, W.L. Chen, R.H. Wang, M.C. Kao, Y.R. Pan, S.H. Chan, K.W. Tsai, H.J. Kung, K.T. Lin, L.H. Wang, Dysregulation of cystathionine gamma-lyase promotes prostate cancer progression and metastasis, EMBO Rep 20 (2019) e45986.
[57] Y. Li, Y. Wang, Y. Zhou, J. Li, K. Chen, L. Zhang, M. Deng, S. Deng, P. Li, B. Xu, Cooperative effect of chidamide and chemotherapeutic drugs induce apoptosis by DNA damage accumulation and repair defects in acute myeloid leukemia stem and progenitor cells, Clin Epigenetics 9 (2017) 83.
[58] P.S. Ward, C.B.J.C.c. Thompson, Metabolic reprogramming: a cancer hallmark even warburg did not anticipate, Cancer Cell 21 (2012) 297-308.
[59] P.P. Hsu, D.M.J.C. Sabatini, Cancer cell metabolism: Warburg and beyond, Cell 134 (2008) 703-707.
[60] D. Anastasiou, Y. Yu, W.J. Israelsen, J.-K. Jiang, M.B. Boxer, B.S. Hong, W. Tempel, S. Dimov, M. Shen, A.J.N.c.b. Jha, Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis, Nature chemical biology 8 (2012) 839-847.
[61] Z. Li, P. Yang, Z. Li, The multifaceted regulation and functions of PKM2 in tumor progression, Biochim Biophys Acta 1846 (2014) 285-296.
[62] K. Zahra, T. Dey, Ashish, S.P. Mishra, U. Pandey, Pyruvate Kinase M2 and Cancer: The Role of PKM2 in Promoting Tumorigenesis, Front Oncol 10 (2020) 159.
[63] M.P.J.B.j. Murphy, How mitochondria produce reactive oxygen species, Biochem. J. 417 (2009) 1-13.
[64] G.A. Spoden, U. Rostek, S. Lechner, M. Mitterberger, S. Mazurek, W.J.E.c.r. Zwerschke, Pyruvate kinase isoenzyme M2 is a glycolytic sensor differentially regulating cell proliferation, cell size and apoptotic cell death dependent on glucose supply, Exp. Cell Res. 315 (2009) 2765-2774.
[65] E. Eigenbrodt, H.J.T.i.p.s. Glossmann, Glycolysis—one of the keys to cancer?, Trends Pharmacol. Sci. 1 (1980) 240-245.
[66] T. Hitosugi, S. Kang, M.G. Vander Heiden, T.W. Chung, S. Elf, K. Lythgoe, S. Dong, S. Lonial, X. Wang, G.Z. Chen, J. Xie, T.L. Gu, R.D. Polakiewicz, J.L. Roesel, T.J. Boggon, F.R. Khuri, D.G. Gilliland, L.C. Cantley, J. Kaufman, J. Chen, Tyrosine phosphorylation inhibits PKM2 to promote the Warburg effect and tumor growth, Sci Signal 2 (2009) ra73.
[67] Z. Yu, X. Zhao, L. Huang, T. Zhang, F. Yang, L. Xie, S. Song, P. Miao, L. Zhao, X. Sun, J. Liu, G. Huang, Proviral insertion in murine lymphomas 2 (PIM2) oncogene phosphorylates pyruvate kinase M2 (PKM2) and promotes glycolysis in cancer cells, J Biol Chem 288 (2013) 35406-35416.
[68] V. Iansante, P.M. Choy, S.W. Fung, Y. Liu, J.G. Chai, J. Dyson, A. Del Rio, C. D'Santos, R. Williams, S. Chokshi, R.A. Anders, C. Bubici, S. Papa, PARP14 promotes the Warburg effect in hepatocellular carcinoma by inhibiting JNK1-dependent PKM2 phosphorylation and activation, Nat Commun 6 (2015) 7882.
[69] G.Y. Liou, P. Storz, Reactive oxygen species in cancer, Free Radic Res 44 (2010) 479-496.
[70] S. Galadari, A. Rahman, S. Pallichankandy, F. Thayyullathil, Reactive oxygen species and cancer paradox: To promote or to suppress?, Free Radic Biol Med 104 (2017) 144-164.
[71] D. Trachootham, J. Alexandre, P. Huang, Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach?, Nat Rev Drug Discov 8 (2009) 579-591.
[72] P. Vaupel, Metabolic microenvironment of tumor cells: a key factor in malignant progression, Exp Oncol 32 (2010) 125-127.
[73] N. Wong, J. Yan, D. Ojo, J. De Melo, J.C. Cutz, D. Tang, Changes in PKM2 associate with prostate cancer progression, Cancer Invest 32 (2014) 330-338.
[74] C. Zhan, Y. Shi, C. Lu, Q.J.D.o.t.E. Wang, Pyruvate kinase M2 is highly correlated with the differentiation and the prognosis of esophageal squamous cell cancer, Dis. Esophagus 26 (2013) 746-753.
[75] C.F. Zhou, X.B. Li, H. Sun, B. Zhang, Y.S. Han, Y. Jiang, Q.L. Zhuang, J. Fang, G.H.J.I.l. Wu, Pyruvate kinase type M2 is upregulated in colorectal cancer and promotes proliferation and migration of colon cancer cells, IUBMB life 64 (2012) 775-782.
[76] S.L. Warner, K.J. Carpenter, D.J. Bearss, Activators of PKM2 in cancer metabolism, Future Med Chem 6 (2014) 1167-1178.
[77] G. Melillo, Targeting hypoxia cell signaling for cancer therapy, Cancer Metastasis Rev 26 (2007) 341-352.
[78] G.L. Semenza, Defining the role of hypoxia-inducible factor 1 in cancer biology and therapeutics, Oncogene 29 (2010) 625-634.
[79] Y. Ju, L. Wu, G.J.B. Yang, b. reports, Thioredoxin 1 regulation of protein S-desulfhydration, Biochemistry 5 (2016) 27-34.